1887

Abstract

The two protein components, II and III, of the 4-chlorobenzoate dehalogenase from sp. CBS3 were cloned separately into . Component II was obtained on plasmid pCBSII, containing a 3.0 kbp dIII fragment, and component III on plasmid pCBSIIIb, containing a 1.3 kbp I/I fragment. The identities of the two components were confirmed by comparison with the authentic components from sp. CBS3. Both components were expressed constitutively in . Neither component alone showed dehalogenating activity. Only in the mixture of crude extracts from both clones was 4-chlorobenzoate dehalogenase detectable. The specific activities in crude extracts were 2.9 mU (mg protein) for component II and 3.5 mU (mg protein) for component III. Expression analysis by minicell experiments revealed a single polypeptide chain of 29 kDa for component II and of 31 kDa for component III.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-137-3-477
1991-03-01
2021-08-06
Loading full text...

Full text loading...

/deliver/fulltext/micro/137/3/mic-137-3-477.html?itemId=/content/journal/micro/10.1099/00221287-137-3-477&mimeType=html&fmt=ahah

References

  1. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  2. Dougan G., Sherratt D. 1977; The transposon Tnl as a probe for studying ColE1 structure and function. Molecular and General Genetics 151:151–160
    [Google Scholar]
  3. Elsner A., Löffler F., Miyashita K., Müller R., Lingens F. 1991; Resolution of 4-chlorobenzoate dehalogenase from Pseudomonas sp. strain CBS3 into three components. Applied and Environmental Microbiology 57: in the Press
    [Google Scholar]
  4. Furukawa K., Tonumura K., Kamibayashi A. 1978; Effect of chlorine substitution on the biodegradability of polychlorinated biphenyls. Applied and Environmental Microbiology 35:223–227
    [Google Scholar]
  5. Gibson T. J. 1984; Studies on the Epstein–Barr virus genome. PhD Thesis University of Cambridge; UK:
    [Google Scholar]
  6. Holmes D. S., Quigley M. 1981; A rapid boiling method for the preparation of bacterial plasmids. Analytical Biochemistry 114:193–197
    [Google Scholar]
  7. Klages U., Lingens F. 1979; Degradation of 4-chlorobenzoic acid by a Pseudomonas spec. Zentralblatt fiir Bakteriologie, Parasitenkunde, Infektionskunde und Hygiene Abteilung Originale CI215–223
    [Google Scholar]
  8. Klages U., Lingens F. 1980; Degradation of 4-chlorobenzoic acid by a Nocardia species. FEMS Microbiology Letters 6:201–203
    [Google Scholar]
  9. Köcher H., Lingens F., Koch W. 1976; Untersuchungen zum Abbau des Herbizids Chlorphenpropmethyl in Boden und durch Mikroorganismen. Weed Research 16:93–100
    [Google Scholar]
  10. Mandel M., Higa A. 1970; Calcium dependent bacteriophage DNA infection. Journal of Molecular Biology 53:159–162
    [Google Scholar]
  11. Maniatis T., Fritsch E. F., Sambrook J. 1982; Molecular Cloning, a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  12. Marks T. S., Smith A. R. W., Quirk A. V. 1984a; Degradation of 4-chlorobenzoic acid by Arthrobacter spec. Applied and Environmental Microbiology 48:1020–1025
    [Google Scholar]
  13. Marks T. S., Wait R., Smith A. R. W., Quirk A. V. 1984b; The origin of the oxygen incorporated during the dehalogenation/hydroxylation of 4-chlorobenzoate by an Arthrobacter sp. Biochemical and Biophysical Research Communications 124:669–674
    [Google Scholar]
  14. Müller R., Thiele J., Lingens F. 1984; Incorporation of 180 water into 4-hydroxybenzoic acid in the reaction of 4-chlorobenzoate dehalogenase from Pseudomonas sp. CBS3. Biochemical and Biophysical Research Communications 124:178–182
    [Google Scholar]
  15. Ruisinger S., Klages U., Lingens F. 1976; Degradation of 4-chlorobenzoic acid by an Arthrobacter species. Archives of Microbiology 110:253–256
    [Google Scholar]
  16. Savard P., Peloquin L., Sylvestre M. 1986; Cloning of Pseudomonas sp. strain CBS3 genes specifiying dehalogenation of 4-chlorobenzoate. Journal of Bacterioloay 168:81–85
    [Google Scholar]
  17. Schneider J., Müller L. 1988; DNA-DNA-Hybridisierung mit biotinylierten DNA-Sonden. Forum Mikrobiologie 6:254–262
    [Google Scholar]
  18. Southern E. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  19. Tautz D., Renz M. 1983; An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels. Analytical Biochemistry 132:14–19
    [Google Scholar]
  20. Thiele J., Müller R., Lingens F. 1987; Initial characterisation of 4-chlorobenzoate dehalogenase from Pseudomonas sp. CBS3. FEMS Microbiology Letters 41:115–119
    [Google Scholar]
  21. Van Den Tweel W. J. J., Ter Burg N., Kok J. B., De Bont J. A. M. 1986; Biotransformation of 4-hydroxybenzoate from 4-chloro-benzoate by Alcaligenes denitrificans NTB-1. Applied Microbiology and Biotechnology 25:289–294
    [Google Scholar]
  22. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
    [Google Scholar]
  23. Zaitsev G. M., Karasevich Yu. N. 1986; Preparative metabolism of 4-chlorobenzoic acid in Arthrobacter globiformis. Mikrobiologiya 50:423–428
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-3-477
Loading
/content/journal/micro/10.1099/00221287-137-3-477
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error